The brown adipose tissue (BAT) is responsible for nonshivering thermogenesis, exhibiting changes in cell morphology related to the functional conditions. In stimulated BAT (neonatal period, cold acclimation) the large majority of adipocytes become multilocular and active, while in inactive BAT (warm acclimation) most cells are pauci- or unilocular and inactive, very similar to white adipocytes. It is well known that white adipocytes are sites of concentration of S-100, a calcium-binding protein originally isolated from the nervous system and later detected also in nonneural cell types, whereas data on the possible presence of this protein in BAT are lacking. The present study used morphological, immunocytochemical, and immunochemical methods to investigate the presence of S-100 protein in BAT under different functional conditions. We found that S-100 was present in both stimulated and inactive BAT and that is was expressed in significantly higher quantities in the latter than in the former. The multilocular cells were always negative (in both active and inactive tissue), whereas the pauci- and unilocular cells were always S-100-positive under both functional conditions. These data suggest that only pauci- and unilocular brown adipocytes express S-100 protein, thus manifesting a possible relationship between S-100 and cell morphology.